Microstructure synthesis by flow lithography and polymerization

ABSTRACT

In a method for synthesizing polymeric microstructures, a monomer stream is flowed, at a selected flow rate, through a fluidic channel. At least one shaped pulse of illumination is projected to the monomer stream, defining in the monomer stream a shape of at least one microstructure corresponding to the illumination pulse shape while polymerizing that microstructure shape in the monomer stream by the illumination pulse. An article of manufacture includes a non-spheroidal polymeric microstructure that has a plurality of distinct material regions.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.60/730,052, filed Oct. 25, 2005, the entirety of which is herebyincorporated by reference.

This application claims benefit as a Divisional Continuation ofapplication Ser. No. 11/586,197, filed Oct. 25, 2006 under 35 U.S.C.§121.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

This invention was made with Government support under Contract No.CTS-0304128 awarded by NSF. The Government has certain rights in theinvention.

BACKGROUND OF INVENTION

This invention relates generally to polymer materials, and moreparticularly relates to techniques for synthesizing polymericmicrostructures.

Polymeric microstructures are important for a wide range ofapplications, including MEMS, biomaterials, drug delivery,self-assembly, and other applications. The ability to controllablysynthesize such microstructures, herein defined as structures havingfeatures in the size range of about 10 nm to about 1000 μm, isincreasingly significant for enabling applications such as paints,rheological fluids, catalysis, diagnostics, and photonic materials.Monodisperse polymeric microstructures, herein defined as having amicrostructure size distribution where >90% of the distribution lieswithin 5% of the median microstructure size, are particularly desirableas they can exhibit a constant and predictable response to externalfields and can self-assemble in a predictable manner.

Conventionally, polymer microstructure synthesis is carried out by abatch process such as photolithography, stamping, or emulsionpolymerization, or by an emulsion-based microfluidic technique such asflow-through microfluidic synthesis. Although these techniques haveprovided significant advances in microstructure synthesis, it is foundin general that each limits microstructure composition and/or geometry.For example, photolithographic techniques generally limit themicrostructure material to that which is compatible with aphotolithographic process, e.g., requiring a photoresist as thestructural material. Historically, the synthesis of polymericmicrostructures with microfluidics has focused almost exclusively onspheroidal microstructures, in part because the minimization ofmicrostructure interfacial energy leads to the formation of spheres ordeformations of spheres such as rods, ellipsoids or discs, or cylinders.

In addition to these limitations in polymeric microstructure compositionand geometry, conventional polymeric microstructure synthesis generallyrequires isotropic structural arrangements of materials. Further, thethrough-put of such processes is typically limited by a requirement formaking one structure at a time or a limited photo-mask-defined, field ofstructures at a time. These limitations in polymeric microstructuresynthesis through-put, microstructure geometry, morphology, andfunctionality have restricted the ability to address the growing numberof critical applications for which polymeric microstructures could bewell suited.

SUMMARY OF THE INVENTION

The invention overcomes the limitations of conventional polymericmicrostructure synthesis to provide lithographic-based microfluidicmicrostructure synthesis techniques that can continuously synthesizepolymeric microstructures of varied complex shapes and chemistries. Inone example polymeric microstructure synthesis method of the invention,a monomer stream is flowed at a selected flow rate through a fluidicchannel; and at least one shaped pulse of illumination is projected tothe monomer stream. This illumination projection defines in the monomerstream a shape of at least one microstructure corresponding to theillumination pulse shape while polymerizing that microstructure shape inthe monomer stream by the illumination pulse.

In a further example synthesis process provided by the invention, amonomer stream is flowed at a selected flow rate, through a fluidicchannel and illumination is projected to the monomer stream topolymerize at least one microstructure in the monomer stream by theillumination. At least one polymerization termination species isprovided, at internal walls of the fluidic channel, which terminates atthe channel walls active polymerization sites at which polymerizationcould occur during polymerization of the microstructure. This quenchespolymerization at those sites and preserves a non-polymerized volume ofthe monomer stream adjacent to the channel walls.

This high-throughput technique enables superior control overmicrostructure geometry, shape, composition, and anisotropy.Non-spheroidal polymeric microstructures each having a plurality ofdistinct material regions can be synthesized by the technique, as canplanar polymeric microstructures each having a plurality of distinctmaterial regions. Other features and advantages of the invention will beapparent from the following description and accompanying figures, andfrom the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A is a schematic view of an example polymeric microstructuresynthesis system in accordance with the invention;

FIG. 1B is a schematic view of the microstructure synthesis system ofFIG. 1A configured with multiple sources of illumination in accordancewith the invention;

FIG. 2 is a schematic cross-sectional view of three polymericmicrostructures as-synthesized in the microstructure synthesis system ofFIG. 1;

FIGS. 3A-3P are schematic views of example polymeric microstructures andthe corresponding lithographic mask employed to produce eachmicrostructure, in accordance with the invention;

FIGS. 4A-4B are schematic views of example chain-like polymericmicrostructures and the corresponding lithographic mask employed toproduce each microstructure, in accordance with the invention;

FIGS. 4C-4D are schematic views of sequential lithography-polymerizationsteps in the synthesis of chain-like polymeric microstructures, and thecorresponding lithographic mask employed to produce each microstructure,in accordance with the invention;

FIG. 5 is a schematic view of a polymeric microstructure includingthree-dimensional features, and the corresponding lithographic maskemployed to produce each microstructure, in accordance with theinvention;

FIGS. 6A-6E are schematic cross-sectional views of five examplemicrofluidic device cross sections that can be employed in accordancewith the invention;

FIG. 7 is a block diagram of a stop flow polymeric microstructuresynthesis control system in accordance with the invention;

FIGS. 8A-8C are schematic views of example polymeric microstructuresincluding selected moieties, and the corresponding lithographic maskemployed to produce each microstructure, in accordance with theinvention;

FIGS. 9A and 9C are schematic views of example flow structures forenabling the polymeric microstructure synthesis in accordance with theinvention across multiple distinct monomer streams;

FIGS. 9B and 9D are schematic views of example polymeric microstructuresincluding multiple distinct polymeric regions produced by thearrangement of FIGS. 9A and 9C, respectively, in accordance with theinvention;

FIGS. 10A-10H are schematic views of example polymeric microstructuresincluding multiple distinct polymeric regions, and the correspondinglithographic mask employed to produce each microstructure, in accordancewith the invention;

FIGS. 11A-11B are plots of measured polymeric microstructure featuresize as a function of illumination exposure duration for threemicrofluidic device channel heights in the system of FIG. 1A and for a20× and 40× microscope objective, respectively;

FIG. 12 is a plot of measured minimum feature size as a function formonomer stream velocity for three microfluidic device channel heights;

FIG. 13 is a schematic view of a flow structure enabling polymericmicrostructure synthesis in accordance with the invention across ahydrophilic monomer stream and a hydrophobic monomer stream;

FIG. 14 is a schematic view of an amphiphilic polymeric microstructure,synthesized by the flow structure of FIG. 13, here defining geometricparameters of the microstructure;

FIGS. 15A-15E are plots of polymeric microstructures counted as afunction of measurement for each of the geometric parameters defined inFIG. 14;

FIG. 16 is a schematic perspective view of an amphiphilic polymericmicrostructure, synthesized by the flow structure of FIG. 13, heredefining geometric parameters that set the radius of curvature of theinterface between a hydrophobic phase and a hydrophilic phase of themicrostructure; and

FIGS. 17A-17B are FTIR plots of transmittance as a function ofwavenumber for three illumination exposure times and for a hydrophobicoligomer and a hydrophilic oligomer, respectively.

DETAILED DESCRIPTION OF THE INVENTION

Referring to FIG. 1A there is shown a schematic view of an examplepolymeric microstructure synthesis system 10 in accordance with theinvention. The system 10 includes a microfluidic device 12 having aselected hollow cross-sectional geometry. The example microfluidicdevice of FIG. 1A exhibits a rectangular channel-like cross section buta wide range of other cross section geometries can be employed asdescribed below. In one example implementation, the microfluidic devicechannel width is characterized by an inner channel width of, e.g.,between about 100 nm and about 1 mm, with channel walls having athickness of, e.g., between about 100 μm and 1 cm, and with a channellength of, e.g., between about 1 mm and several centimeters. Themicrofluidic device is configured to accept a monomer stream 15 that isdirected to the hollow cross section, or channel, of the microfluidicdevice for passage through the device. The microfluidic device is formedof any suitable material, as explained below. In the example arrangementof FIG. 1A the microfluidic device can be constructed of, e.g.,polydimethylsiloxane (PDMS) or other suitable material.

The monomer stream 15 can include a range of constituents, as explainedin detail below. At least one of the constituents is provided as aliquid-phase monomer that can be polymerized by a selectedpolymerization process, e.g., photo-polymerization, thermalpolymerization, or other process. In the example system of FIG. 1A themonomer stream includes a photo-polymerizable monomer and aphotosensitive initiator species. One example of a suitablephoto-polymerizable monomer is poly(ethylene glycol) diacrylate(PEG-DA), e.g., having a molecular weight of 400, with2-hydroxy-2-methyl-1-phenyl-propan-1-one employed as the photosensitiveinitiator species. The monomer stream can also include other selectedmonomers, as well as particles, molecules, porogens, and other speciesas explained below. Whatever its composition, the monomer streamincludes at least one liquid-phase polymerizable monomer that enablespassage of the stream through the microfluidic device 12.

At one or more points along the microfluidic device is providedstimulation for enabling the formation and polymerization ofmicrostructures in the monomer stream. For the example of FIG. 1A inwhich the monomer stream includes a photo-polymerizable monomer, at oneor more selected points along the length of the microfluidic devicethere is provided a source 16 of illumination 17 that is directed towardthe microfluidic device 12. The walls of the microfluidic device arepreferably substantially transparent to the wavelength of theillumination 17. Visible light, UV light, IR light, or other wavelengthof light can be employed as-suited for a selected monomer species, asexplained in detail below. For the PEG-DA monomer described above, UVillumination is a suitable polymerizing radiation.

The polymerizing radiation is shaped in correspondence with desiredpolymeric microstructure shapes. For example, interposed between theillumination source and the microfluidic device can be provided one ormore lithographic masks or other lithographic system for shaping theillumination. In the example of FIG. 1A, there is provided a dark fieldlithographic mask 18 including one or more shapes 20 desired forpolymeric microstructures to be synthesized. As shown in FIG. 1A themask can include a plurality of distinct shapes or can include a numberof replications of a single shape.

A lens system 22 can be interposed between the lithographic mask and themicrofluidic device if desired for controlling magnification, focus, orother aspect of the illumination 17 directed through the mask. Theillumination exits the lens system and is directed to the microfluidicdevice. In accordance with the invention, the illumination is temporallycontrolled to provide pulses of illumination of a selected duration. Ashutter 23 or other mechanism for controlling the duration ofillumination is preferably provided in a suitable configuration with thelens system 22 and illumination source 16. The duration of eachillumination pulse is set based on the flow rate of the monomer stream,the polymerizing characteristics of the monomers in the stream, and thedesired shape of a microstructure. As explained in detail below, theflow rate of the monomer stream is also controlled, and can be stopped,in coordination with the temporal control of the illumination. Theillumination pulses can be provided as a sequence of pulses, each of aselected duration, or as a single long-duration pulse, as-prescribed fora given application.

Referring also to FIG. 1B, there can be provided multiple illuminationsources and lens systems 22, 35, 37, along the length of themicrofluidic device and/or at locations around selected walls of thedevice, for delivering pulses of illumination to a monomer stream fromdifferent angles and locations around the microfluidic device. Forclarity in FIG. 1B there are shown lens systems 22, 35, 37, but such areintended to represent the inclusion of shuttering systems, illuminationsources, and illumination masks or other shaping devices in conjunctionwith the lens systems as in the manner of FIG. 1A and described justabove.

Referring now back to FIG. 1A, exposure of the monomer stream 15 passingthrough the microfluidic device 12 to a pulse of the shaped illumination24 polymerizes mask-defined microstructure shapes 30 directly in themonomer stream. The illumination exposure simultaneously defines theshapes of polymeric microstructures and polymerizes the shapedmicrostructures. This dual lithography-polymerization action occurs inthe continuous phase of the monomer stream; that is, the one or moreliquid-phase polymerizable monomers in the stream operate as acontinuous phase of the stream and are themselves polymerized. Thus thepolymerized microstructures resulting from the duallithography-polymerization action include polymeric material from thecontinuous phase of the monomer stream.

As stated above, flow of the monomer stream can be controlled in acoordinated manner with the illumination exposure to in turn controlcharacteristics of the microstructure polymerization. The flow rate andexposure duration are preferably together selected such that there issufficient dwell time of a given volume of the monomer stream at thesite of illumination exposure for substantially full polymerization ofmask-defined microstructures in the stream. If desired, the monomerstream flow can be substantially stopped in coordination withillumination pulse exposure. With continuous monomer stream flow, a highsynthesis through-put, e.g., 100 microstructures per second, can beachieved. Such can further be enhanced with the inclusion of multipleillumination points along the length of the microfluidic device, in themanner described above, and by increasing the illumination area and thecorresponding number of microstructure shapes projected to the increasedarea.

As shown in FIG. 1A, once polymeric microstructures 30 are polymerizedin the monomer stream, the polymerized microstructures advect throughun-polymerized monomer of the stream through the microfluidic device. Aset 32 of such polymerized structures are schematically shown in FIG. 1Adownstream of the lithography-polymerization point. The volume ofun-polymerized continuous-phase monomer remaining in the stream afterthe lithography-polymerization step operates to conduct the polymerizedmicrostructures through and out of the microfluidic device. A reservoir34 is provided for collecting the output monomer stream 36, whichincludes a population 38 of polymerized microstructures. As can berecognized, if only one transparent mask shape is employed, then themicrostructure population is homogeneous, and preferably ismonodisperse.

The synthesized microstructures can be rinsed in the reservoir or, e.g.,pipetted into another container for rinsing. For example, the monomerstream including the microstructure population can be pipetted from thereservoir into an eppendorf tube, suspended in a buffer with asurfactant to prohibit agglomeration, and centrifuged to retrieve themicrostructure population from the monomer stream. The microstructurepopulation can then be employed for a selected application.

The ability of the monomer stream to conduct the polymerizedmicrostructures through and out of the microfluidic device is enabled inaccordance with the invention by preserving a non-polymerized volume ofthe stream at the location of the walls of the microfluidic device aswell by the unpolymerized stream volume between microstructures, atlocations that were masked from the polymerizing illumination.Preservation of a non-polymerized volume of the stream at the locationof the device walls prohibits polymerization from extending to the wallsof the device and inhibits adhesion of polymerized microstructures tothe device walls. In general, in accordance with the invention, this isachieved by providing at the internal faces of the microfluidic devicewalls a species that can participate in a polymerization terminationstep; i.e., a species that can react to terminate active polymerizationsites at which polymerization could occur, such that polymerization isquenched at those sites near to the device walls.

The selected polymerization termination species can be provided at theinternal walls of the microfluidic device in any convenient mannersuited for a given application. For example, the polymerizationtermination species can be directed to pass through the hollowcross-sectional channel of the microfluidic device at the walls of thedevice, as described in more detail below. Alternatively, thepolymerization termination species can be provided by diffusion of thespecies through the microfluidic device walls from the ambient to theinternal wall faces. For many applications, this arrangement can beparticularly convenient and elegantly simple to implement; all that isrequired is that the microfluidic device be provided as a material thatis sufficiently porous to enable diffusion of the selected terminationspecies while sufficiently solid to contain the monomer stream in thedevice.

In one example of polymerization termination species diffusion throughthe microfluidic device walls, the termination species is specified asambient oxygen and the monomer to be polymerized is specified as afree-radical-polymerization monomer, e.g., the PEG-DA monomer describedabove, with the 2-hydroxy-2-methyl-1-phenyl-propan-1-one photoinitiatordescribed above. The microfluidic device is here formed of PDMS, asdescribed above, which is porous to oxygen, or is formed of anothersuitable oxygen-permeable material, such as Mylar, polyurethane,polyethelene, polychlorophene, mercapto ester-based resins, e.g.,Norland 60, from Norland Optical Products, Inc., New Brunswick, N.J.,porous Tygon® tubing from Saint-Gobain Performance Plastics, Mickleton,N.J., or other material.

When a PEG-DA-based monomer stream passing through the PDMS device isexposed to a pulse of UV illumination, the2-hydroxy-2-methyl-1-phenyl-propan-1-one photoinitiator forms freeradicals, some of which are passed to the PEG-DA monomer for initiatingpolymerization of the PEG-DA monomer. Oxygen diffusing through the PDMSwalls of the microfluidic device reacts with these free radicals toterminate the molecular free-radical sites, both on the initiatorspecies and on the monomer. The free-radical sites are thereby convertedto chain-terminating peroxide radicals. Polymer chain growth is theninhibited, or quenched, at the converted sites.

Near to the internal faces of the PDMS device walls, this polymerizationtermination process continues as oxygen is consumed in thechain-terminating conversion of free-radical sites in the monomer streamnear to the device walls, causing more oxygen to diffuse through thedevice walls. This results in the formation of a thin un-crosslinked,non-polymerized lubricating monomer layer at the microfluidic devicewalls. The lubricating monomer layer provides a volume ofcontinuous-phase monomer stream through which polymerizedmicrostructures can advect and prohibits adhesion of the microstructuresto the device walls.

With this oxygen-aided polymerization inhibition technique, theinvention provides a discovery that free-radical polymerizationtermination, which is typically considered to be a hindrance for mostapplications, can be exploited and controllably enforced to enablepolymerization directly in a continuous phase monomer stream withpolymerization inhibited at edges of the monomer stream andnon-polymerized monomer preserved for transporting polymerizedmicrostructures dispersed in the monomer steam. The example oxygen-basedtermination process is applicable to any free-radical polymerizationsystem. For example, monomers such as 1,1,1-tri(methyl propanetriacrylate), 1,6-hexanediol diacrylate, and poly(ethylene glycol)dimethacrylate, among others, as identified in detail below, can beemployed, with photoinitiators such as DMPA or IRGACURE 184(1-Hydroxy-cyclohexyl-phenyl-ketone), from Ciba Specialty Chemicals,Tarrytown, N.Y.

The thickness of the non-polymerized lubricating monomer layer at themicrofluidic device walls is set by the concentration of species in themonomer stream for which free radicals are generated by the lithographicillumination, by the local intensity of the illumination, and by thedegree of terminating species, e.g., oxygen flux, through the devicewalls. In addition, it is to be recognized that each polymerizationchemistry is characterized by distinctive reaction kinetics that adjustfree radical concentration in a monomer stream. Thus, for manyapplications, empirical analysis can be preferred to determine processparameters that provide a sufficient lubricating monomer layer. For manyapplications, a lubricating layer thickness of between about 1 μm andabout 5 μm is sufficient to prohibit microstructure adhesion to walls asthe microstructures are carried out of the microfluidic device.

The polymerization termination process of the invention can beimplemented with a range of polymer chemistries and terminating species.For example, free-radical terminating species such as HQ (hydroquinone)or MEHQ (monomethyl ether hydroquinone) and suitably permeable devicematerials can be used to terminate free-radical species in themicrostructure polymerization process. In addition, a range offree-radical polymerization-initiating species can be employed in apolymerization that is not necessarily photo-initiated. For example,thermally-activated free-radical initiators, e.g.2,2′-Azobis(2-methylpropionitrile), can be used with an appropriate heatsource, e.g. laser illumination or patterned conducting electrodes, tocreate free-radicals for free-radical-based polymerization; oxygen orother terminating species being employed to prohibit polymerization atthe microfluidic device walls.

The process is further not limited to free-radical termination. Forexample, given the photoresist SU-8, which is based on EPON SU-8 epoxyfrom Shell Chemical, NY, N.Y., as a polymerizing species in the monomerstream, termination of SU-8 polymerization at the microfluidic devicesidewalls can be implemented with a selected base species, such as3-ethylamine, 3-octylamine, or other suitable base that quenches SU-8polymerization. The selected base species can be provided to the monomerstream at internal faces of the microfluidic device sidewalls bydiffusion through the sidewalls, given a suitable porosity in thesidewalls. For example, a device of PDMS or other suitable porousmaterial can be employed for diffusing a liquid base or other liquidspecies through the device. Here the device can be initially saturatedwith the selected liquid species, with the liquid continuously suppliedto the outer surface of the device, e.g., in a bath arrangement, toreplenish the liquid as it is consumed within the microfluidic device.

Given an arrangement in which a terminating species is supplied tointernal faces of the microfluidic device walls by diffusion through thewalls, the material of the microfluidic device is preferablysufficiently permeable to the terminating species of interest. Forexample, for the terminating species oxygen, the device walls preferablyhave an O₂ gas permeability of at least about 10 barrer. For theterminating species of a selected liquid, the device walls preferablyhave a liquid permeability of about 10 barrer.

The microfluidic device can be polymeric, as in the example materialsdescribed above, or can be non-polymeric. For example, glassmicrofluidic device walls, with suitable nano-scale holes forpermeability of a selected terminating species, can be employed.Microfluidic device walls can also be provided as, e.g., track-etchedmembranes, and other structures, including, e.g., porous silicasubstrates or other inorganic structures, such as glass slides. It isnot required that all walls be permeable, but it is preferred that thosewalls exposed to a polymerizing agent, such as illumination, bepermeable to a selected terminating species for enabling termination ofpolymerization at those walls. Of course, as explained above, the one ormore walls through which illumination is directed to a monomer streamare preferably substantially transparent to the illumination wavelength,and illumination can be directed through multiple device walls forinducing polymerization termination at those walls.

In accordance with the invention, a terminating species can also beintroduced into the monomer stream flow channel along with the monomerstream instead of diffusing into the channel through the microfluidicdevice walls. In one example arrangement of such, an annular sheath flowof fluid containing a selected terminating species is provided at themicrofluidic device channel inlet to enclose an inner cylindrical flowof the monomer stream. In another example arrangement, two fluids withdifferent wetting properties are introduced at a Y-junction input to thechannel, in a manner discussed in more detail below, such that thefluids flow parallel to each other. Process conditions can here be tunedsuch that the fluid containing the terminating species preferentiallywets the surface walls of the device, leading to an enveloping of themonomer stream species to be polymerized.

Referring now also to FIG. 2, which is a cross-sectional view of themicrofluidic device 12 in FIG. 1A taken at the point along the device inwhich shaped illumination 24 is projected to the device, the shapes ofthe polymerized microstructures 30 in the x-y plane, defined in FIG. 1A,are determined by the shapes of the features 20 included on thetransparency mask 18. The z-plane projection of the microstructures isdependent on the height of the microfluidic device cross-section and thethickness of the polymerization inhibition layers 40 produced at thosewalls that are exposed to the illumination. The polymerizationinhibition layer thickness is independent of device height, and thus hasa more-pronounced effect on microstructures synthesized in low-heightdevices, where the layer occupies a larger fraction of the deviceheight.

FIGS. 3A-3G present a range of example polymeric microstructureconfigurations that are enabled by the invention with a synthesis systemlike that of FIG. 1. Each of FIGS. 3A-3G schematically represent anexample shape 50 of a transparency mask and the corresponding polymericmicrostructure 52 produced by the synthesis process described above. Asshown in these figures, curved, straight-edged, hollow geometries, in anearly arbitrary combination, can be produced. Many of themicrostructures can be characterized as planar microstructures in thattheir extent in a selected plane, e.g., the x-y plane, is substantiallylarger than that in the z-plane. Referring also to FIGS. 3H-3J, if theshape is produced with a gray-scale mask configuration 55, wheregray-scale features of a shape are provided on the mask, thencorresponding shaped-profile microstructures 58 result from themicrostructure synthesis process of the invention, with tailoring of themicrostructure profile in the z-plane.

Referring now to FIGS. 3K, 3L, 3M, and 3N, ribbon-like structures canalso be produced by the lithography-polymerization step. As shown inFIG. 3K, a selected mask 60 is employed to set the width and otherfeatures of the ribbon microstructure 62. The mask can be, e.g.,rectangular, square, round, or other geometry to set the widthcharacteristic of the extruded ribbon structure. In formation of such aribbon-like structure, the projection of illumination is not pulsed inthe manner described above, and instead is continuously maintained on asthe structure is lithographically defined and polymerized and thentransported through the microfluidic device.

As shown in FIG. 3L, a dynamic mask technique can be employed to imposechangeable mask shapes on a polymerizing ribbon microstructure as themicrostructure is extruded through the fluidic microdevice. As explainedin detail below, such a dynamic mask technique can be implemented by,e.g., a digital micromirror device (DMD) or other suitable device thatenables changes in mask features while continuous illumination ismaintained during the lithography-polymerization process. In the exampleof FIG. 3L, three changing mask shapes 64, 66, 68 are employed toproduce a ribbon structure 70 having a sequence of distinct shapes alongthe length of the ribbon. This technique enables wide flexibility incustomization of ribbon microstructure features.

As shown in FIG. 3M, features of the ribbon structure profile can alsobe adjusted by a dynamic mask technique as the ribbon is polymerized.For example, given two rectangles 72, 74 of differing length, a ribbonstructure 76 having a modulating width can be produced by alternatingexposure of a continuously-flowing monomer stream between the two maskgeometries. Similarly, as shown in FIG. 3N, with a single rectangle 72and two rectangles 74 employed as two distinct mask features, a ribbonstructure 78 can be produced having both modulation in the ribbon widthas well as changes in features along the ribbon.

Referring to FIGS. 3O-3P, there can be produced polymericmicrostructures 77, 79 having conical profiles that result fromconditions of illumination that are controlled in combination with themicrostructure mask shape. As shown in FIG. 3O, a circular mask shape 73is here employed. This circular mask shape can produce the structures ofFIGS. 3O-3P when the microfluidic device channel is sufficiently tallthat points in the channel are out of the depth of field of illuminationthat is focused on the channel. If the illumination is focused on thecenter of the channel and the depth of field is smaller than the heightof the channel, then the double-conical microstructure of FIG. 3P issynthesized. If the illumination is focused at a non-center point of thechannel and the depth of field is smaller than the height of thechannel, then the conical microstructure of FIG. 3O is synthesized, withthe extent of each conical structure is determined by the location ofthe focus relative to the channel height.

Now referring to FIGS. 4A-4B, if the monomer stream flow rate andillumination pulse duty cycle are controlled to cause polymerization atdirectly adjacent or slightly overlapping volumes of the monomer stream,then a mask feature 80, 82 can be employed to form chain-likemicrostructures 84, 86, respectively, where each chain is formed as asequence of connected polymeric shapes.

FIG. 4C schematically represents an example sequence oflithography-polymerization events employed for producing the chainstructure 86 of FIG. 4B. With the monomer stream flow from left to rightas indicated in the figure, pulses of illumination are repeated as themonomer stream continuously flows such that each polymeric shape isappended to the growing chain-like structure, with each additionalpolymeric shape linked by an overlapping polymerized region. Forexample, at a first illumination pulse (1), a first cylindricalstructure is formed; at a second pulse (2) a second cylindricalstructure is appended to the first; then at a third pulse (3) a thirdcylindrical structure is appended to the second; and at a fourth pulse(4) a fourth cylindrical structure is appended to the third. In thismanner the chain-like structure can be extended to a desired length.

FIG. 4D schematically represents an example sequence oflithography-polymerization events employed for producing a chain-likestructure with a dynamic mask technique that enables adjustment orchange of the microstructure shape being appended to a chain-likemicrostructure. Again with the monomer stream flow from left to right asindicated in the figure, pulses of illumination are repeated as themonomer stream continuously flows such that each shape unit is appendedto the growing chain-like structure, with each shape unit linked by anoverlapping polymerized region. At a first illumination pulse (1), afirst cylindrical structure is formed; at a second pulse (2) arectangular structure is appended to the cylinder; at a third pulse (3)a square structure is appended to the rectangle; at a fourth pulse (4) arectangle is appended to the square; and at a fifth pulse (5) a triangleis appended to the rectangle. This dynamic mask changing enables a highdegree of customization of a chain-like structure and can produce anear-arbitrary range of chain combinations.

The shapes and feature geometries of FIGS. 3-4 are representative of thepolymeric microstructures that can be produced in accordance with theinvention, but are not exhaustive. These examples demonstrate that bothdiscrete, particle-like microstructures, as well as interconnectedmicrostructures, such as ribbons or chains, can be produced by themicrostructure synthesis process of the invention. The microstructurescan non-continuous across a plane of the microstructure, as in theexample microstructures of FIGS. 3D-3F. Additional microstructuregeometries can include, e.g., polygonal features, such as hexagons,colloidal cuboid microstructures, high-aspect-ratio objects such asposts with circular, triangular, or square cross sections, andnon-symmetric objects. The term “microstructure” is herein meant torefer to any structure having feature sizes on the length scale of about10 nm to about 1000 μm. No particular geometry or symmetry is implied bythe term. But circular, rounded, symmetric and other such geometries canbe produced in accordance with the invention. For many applications, theterm “microparticle” can also be employed to describe microstructuresthat are distinct, separate entities which can be dispersed in acontinuous phase. The ribbon and chain-like microstructures descriedabove are not for most applications characterized as microparticles, butare microstructures as contemplated by the invention.

The correspondence between a mask shape and a resulting microstructuregeometry is in part based on the lens system employed to implement theprojection lithography of the single-step lithography-polymerizationprocess. For many applications, it can be convenient to employ as thelens system an inverted microscope objective. In this example, the maskfeatures' sizes are reduced by a factor that is dictated by thecharacteristics of the microscope objective and the other lenses in thesystem. Typically an additional lens is in the optical path between thefield-stop slider and the objective of a microscope lens system. Then,for example, given a ×20 objective, a 7.8 times feature reductionresults due to a 2.57× lens between the objective and the field-stopslider. In this case, a 350 μm-square mask feature results in synthesisof microstructure cuboids, i.e., rectangular parallelepiped objects,having 45 μm-long sides in the x-y plane, as defined in FIGS. 1-2.

The height of a microstructure in the z plane, as defined in FIGS. 1-2,is for most two-dimensional microstructures, set by the cross section ofthe microfluidic device and the non-polymerized lubricating layers inthe device. For the oxygen-aided polymerization inhibition processexample given above, a non-polymerized lubricating layer of betweenabout 1 μm and about 5 μm in height is sufficient. With anon-polymerized lubricating layer of about 2.5 μm in thickness, thengiven a microfluidic device cross-sectional height of 20 μm a resultingpolymeric microstructure is about 5 μm shorter, or in this example,about 15 μm tall in the z plane as defined in FIGS. 1-2.

For many applications, the fundamental limitations of a projectionlithographic technique like that of the example described above are setby the optical resolution and depth of field of the lens objectiveemployed. The resolution of an objective herein is meant to refer to thesmallest distinguishable feature that can be discerned, and the depth offield is herein meant to refer to the length over which a beam of lightemanating from an objective can be considered to have a constantdiameter. In the projection lithography technique of the invention,optical resolution limits the minimum feature size that can besynthesized, and the depth of field restricts the length over which thesidewalls of a polymerized microstructure will be straight. An increasein optical resolution decreases the depth of field. In addition, theminimum feature size that can be printed on a transparency maskcontributes to feature size limitations.

If a suitable transparency mask can be produced for a given application,no lens system need be employed in the projection lithography process.In the absence of a lens objective, illumination passes through a maskand is directed to a monomer stream in a microfluidic device without anyreduction in feature size. The mask features are then identicallyreproduced in the polymerizing monomer stream.

The invention is not limited to a particular type of mask, and anysuitable mask or illumination arrangement that enables the simultaneousshape-definition and polymerization of polymeric microstructures in amonomer stream can be employed. Apertures, ink masks, metallic maskssuch as chrome masks, photographic film, dynamic masks such as thedigital micromirror devices described above, and other such masks andtechniques can be employed. In general, all that is required is anability to project or direct illumination toward a monomer stream in amicrofluidic device.

Multiphoton illumination and multiple-beam lithographic techniques canalso be employed, e.g., to produce three-dimensional microstructurefeatures. FIG. 5 is a schematic view of an example polymericmicrostructure 90 synthesized in accordance with the invention andincluding 3-D features in the microstructure. In this example, atriangular mask feature 92 is employed with laser interferencelithography to lithographically define the microstructure as the laserillumination polymerizes the microstructure. The resultingmicrostructure is characterized by a matrix having interpenetratingholes throughout the structure. The added 3-D dimensionality to apolymeric microstructure of the invention can be employed for a range ofapplications for a selected functionality or other action. The inventionis not limited to a particular 3-D technique or class of microstructuresand enables a near-arbitrary 3-D customization of microstructurefeatures.

The cross-sectional geometry of the microfluidic device employed in thesynthesis process of the invention can also be tailored to customizemicrostructure geometry. FIGS. 6A-6E are schematic cross-sectional viewsof example microfluidic device geometries. In these examples, there isprescribed a microfluidic device assembly process that enablesconvenient fabrication of a wide range of device geometries.

In FIG. 6A there is shown an example device 12 like that of FIGS. 1A-1B,having a rectangular cross sectional volume 95 through which a monomerstream can pass. Fabrication of this device can be conveniently carriedout for polymeric device materials, e.g., PDMS. In such a scenario, amold in, e.g., a silicon substrate that is lithographically patternedand etched with a selected device cross sectional geometry, is provided.A corresponding PDMS structure 100 is formed by conventional moldingtechniques with the substrate mold. The resulting PDMS structure 100forms the top and sides of the device. A PDMS layer 102 is provided on atransparent support structure, e.g., a glass plate 104, to provide thebottom of the PDMS device. The PDMS structure 100 is mated with the PDMSlayer 102 on the glass plate to form the hollow rectangular microfluidicdevice. This assembly process is particularly convenient and enablesprovision of a support structure such as a glass plate for the device.

Referring now to the example of FIG. 6B, the top PDMS structure 100 canbe molded as described above, with a patterned and molded PDMS layer 106is provided on a glass plate 104. This enables a cross section, orchannel, 108 having features along the width of the device. As shown inFIG. 6C, this example can be extended by employing multi-levellithographic techniques to pattern both a PDMS structure 110 and a PDMSlayer 112. This combination of lithographic steps enables production ofa complex device cross section 114.

FIG. 6D schematically represents an example microfluidic device crosssection wherein the PDMS structure 116 is molded around a tube or rod toproduce a rounded cross section 118. The molded structure 116 can itselfbe supported directly on, e.g., a glass plate 104. FIG. 6E schematicallyrepresents an example microfluidic device wherein a PDMS structure 120is formed using a grayscale lithographic technique to enable angledgeometries across the width of the structure. An angled cross section122 results from this configuration. Here a PDMS layer 102 can beprovided on a glass plate 104 as described above, or lithographicdefinition of the PDMS layer can also be employed to add to features ofthe cross section profile.

The height of a microfluidic device cross section, in combination withthe feature size of a projection mask, sets the exposure time requiredto fully polymerize microstructures in the photosensitive monomerexample of FIG. 1 described above. The polymerization exposure time isinversely proportional to both the device cross section and mask featuresize. When the extent of the channel in a direction aligned with theprojected illumination is decreased or when the mask feature size isdecreased, an increased polymerization duration is required. A reductionin mask feature size requires an increase in polymerization dose due todiffraction-induced limitations in, the optical train of a microscopeemployed for focusing illumination, or other optical system employed.

An increase in required polymerization duration in turn places aconstraint on the maximum allowable velocity of a continuously-flowingmonomer stream passing through a microfluidic device, to avoidunintended shape deformation of polymerized microstructures in thestream. In accordance with the invention, it can be preferred to specifya microstructure feature size tolerance and based on that tolerance andthe polymerization duration required for the microfluidic devicedimensions and the mask feature size, to determine a monomer stream flowrate that accommodates the specified microstructure feature sizetolerance. The flow rate of the monomer stream can be controlled by,e.g., syringe pump operation, capillary action, pressure, electrokineticforce, or other selected operation.

As described above, stop flow lithography can be employed as analternative to continuous flow lithography in accordance with theinvention with a monomer stream in a microfluidic device like that ofFIG. 1. In a stop flow lithography process, the monomer stream flow isbrought substantially to a complete stop before alithography-polymerization step is initiated. At the completion of thelithography-polymerization step, the monomer flow is resumed and thesynthesized particles are completely flushed out of the region in thechannel in which the lithography-polymerization takes place. Thisensures that synthesized microstructures do not interfere with thepolymerization of subsequent microstructures.

For many applications, the stop flow lithography technique can providesynthesized polymeric microstructure feature resolution that is improvedover that of continuous flow lithography. The stop flow lithographytechnique can also achieve a microstructure synthesis through-put thatis increased over that of continuous flow lithography because a higheraverage monomer stream flow rate can be employed, considering that themicrostructure flushing step can be carried out at a high flow rate.Continuous flow lithography is in general mechanistically simpler thanstop flow lithography and thereby can be preferred for applications inwhich the distinct advantages of stop flow lithography are not required.Stop flow lithography can be preferred to continuous flow lithographyfor applications employed to produce high-fidelity microstructures withsmall features, e.g., ˜10 μm or less, or when it is desired to producesharp interfaces between adjacent chemistries in a microstructure, asdescribed below. The interface between two miscible streams is sharperfor smaller residence times during which molecular species can diffusebetween the adjacent streams. In a stop-flow lithography setup, one canpolymerize immediately after stopping two rapidly-flowing streams tominimize the residence time and increase the sharpness of theinterfaces.

FIG. 7 is a block diagram of the elements of an example stop flowlithography system 150 that can be employed for carrying out thephotosensitive polymerization of microstructures as described with FIG.1 above. The stop flow lithography system 150 includes a controlledpressure source 152 for pressure driven flow of a monomer stream and a3-way valve 154 to stop and start pressure from the source 152. Apressure source can be preferred over a syringe pump for thisapplication due to the better dynamic response of the pressure sourcearrangement.

A light source, e.g., a UV light source 156 is provided for projectionlithography. A shutter 158 or other control mechanism is provided forcontrolling duration of the illumination and a mask 160 or other systemis aligned with a microfluidic device 162 in the manner described above,and a microscope objective 164 or other lens system is provided, ifdesired, for reducing the illumination feature size. A computer 166 orother processing device or system is provided for synchronouslycontrolling the shutter 158 and the valve 154 such that the monomerstream flow through the microfluidic device is coordinated to stop andstart in unison with illumination of the monomer stream.

The valve and shutter can be controlled by way of, e.g., serial RS 232connections, USB connections, through a data acquisition board, or othersuitable connection, with suitable control software, such as LabVIEW,National Instruments Corp, Austin, Tex. Alternatively, acomputer-controlled pressure transducer can be employed to adjust themonomer stream pressure between a specified flowing pressure and zeropressure of stopped flow. The pressure source can be provided as anysuitable gas pressure source that can supply between, e.g., about 0-30psi. The 3-way valve or the pressure transducer is connected to themicrofluidic device through suitable tubing, e.g., through 1/32″diameter Tygon® tubing that is fitted to the device by, e.g., a 10 μlpipette tip. In one example scenario, the pipette tip is filled with avolume of monomer stream and inserted in the microfluidic device forinitiating monomer flow through the device. A compressed gas head canthen be employed to drive the flow by attaching the tubing through whichthe air flows to the pipette tip.

While the system 150 of FIG. 7 has been described specifically for astop flow lithography process, the system 150 can also be employed forcontrolling flow rate of a monomer stream in a continuous flowlithography process. In this mode of operation, the 3-way valve ismaintained open and a constant pressure is applied to the monomer streamflowing through the device. The computer can here be employed forcontrolling monomer stream flow rate as a function of shutter speed.

Turning now to specific features of the polymer chemistry employed formicrostructure synthesis processes of the invention, the processes canbe conducted with any polymerizable liquid-phase monomer in whichmicrostructure shapes can be defined and polymerized in a singlelithography-polymerization step. Preferably the selected monomer also ischaracterized by a polymerization reaction that can be terminated with atermination species. The terminating species, lithographic illumination,and monomer constituents are therefore selected in cooperation to enableall such functionality of the process of the invention.

As explained above, one particularly well-suited class of polymers isthat class of polymerizable monomers that undergo free-radical-initiatedpolymerization, e.g., by UV illumination, visible light illumination,thermal initiation, or other initiating radiation or agent. Such monomersystems preferably include one or more unsaturated (double bond) speciesthat undergo free-radical-initiated polymer chain extension. Examples ofsuch monomers include acrylates, multi-acrylates, methacrylates,multi-methacrylates, vinyls, and any blends thereof.

Table I below is a non-exhaustive listing of a range of monomers thatcan be employed for polymeric microstructure synthesis in accordancewith the invention.

TABLE I Allyl Methacrylate Benzyl Methylacrylate 1,3-ButanediolDimethacrylate 1,4-Butanediol Dimethacrylate Butyl Acrylate n-ButylMethacrylate Diethyleneglycol Diacrylate Diethyleneglycol DimethacrylateEthyl Acrylate Ethyleneglycol Dimethacrylate Ethyl Methacrylate 2-EthylHexyl Acrylate 1,6-Hexanediol Dimethacrylate 4-Hydroxybutyl AcrylateHydroxyethyl Acrylate 2-Hydroxyethyl Methacrylate 2-HydroxypropylAcrylate Isobutyl Methacrylate Lauryl Methacrylate Methacrylic AcidMethyl Acrylate Methyl Methacrylate Monoethylene Glycol2,2,3,3,4,4,5,5-Octafluoropentyl Acrylate Pentaerythritol TriacrylatePolyethylene Glycol (200) Diacrylate Polyethylene Glycol (400)Diacrylate Polyethylene Glycol (600) Diacrylate Polyethylene Glycol(200) Dimethacrylate Polyethylene Glycol (400) DimethacrylatePolyethylene Glycol (600) Dimethacrylate Stearyl MethacrylateTriethylene Glycol Triethylene Glycol Dimethacrylate2,2,2-Trifluoroethyl 2-methylacrylate Trimethylolpropane TriacrylateAcrylamide N,N,-methylene-bisacryl-amide Phenyl acrylate Divinyl benzene

For those monomers that are photo-polymerizable, a photoinitiatorspecies is included in the monomer stream to enable the polymerizationprocess. Effectively any chemical that can produce free-radicals in thefluidic monomer stream as a result of illumination absorption can beemployed as the photoinitiator species. There are in general two classesof photoinitiators. In the first class, the chemical undergoesunimolecular bond cleavage to yield free radicals. Examples of suchphotoinitiators include Benzoin Ethers, Benzil ketals,a-Dialkoxy-acetophenones, a-Amino-alkylphenones, and Acylphosphineoxides. The second class of photoinitiators is characterized by abimolecular reaction where the photoinitiator reacts with a coinitiatorto form free radicals. Examples of such are Benzophenones/amines,Thioxanthones/amines, and Titanocenes (vis light).

Table II below is a non-exhaustive listing of a range of photoinitiatorsthat can be employed with a photo-polymerizable monomer for polymericmicrostructure synthesis in accordance with the invention.

TABLE II Trade Name (CIBA) Chemical Name IRGACURE 1841-Hydroxy-cyclohexyl-phenyl-ketone DAROCUR 11732-Hydroxy-2-methyl-1-phenyl-1-propanone IRGACURE 29592-Hydroxy-1-[4-(2-hydroxyethoxy) phenyl]-2-methyl- 1-propanone DAROCURMBF Methylbenzoylformate IRGACURE 754 oxy-phenyl-acetic acid 2-[2 oxo-2phenyl-acetoxy- ethoxy]-ethyl ester and oxy-phenyl-acetic 2-[2-hydroxy-ethoxy]-ethyl ester IRGACURE 651 Alpha,alpha-dimethoxy-alpha-phenylacetophenone IRGACURE 3692-Benzyl-2-(dimethylamino)-1-[4-(4-morpholinyl) phenyl]-1-butanoneIRGACURE 907 2-Methyl-1-[4-(methylthio)phenyl]-2-(4-morpholinyl)-1-propanone DAROCUR TPO Diphenyl (2,4,6-trimethylbenzoyl) phosphineoxide IRGACURE 819 Phosphine oxide, phenyl bis (BAPO) (2,4,6-trimethylbenzoyl) IRGACURE 784 Bis (eta 5-2,4-cyclopentadien-1-yl) Bis[2,6-difluoro- 3-(1H-pyrrol-1-yl) phenyl]titanium IRGACURE 250 Iodonium,(4-methylphenyl) [4-(2-methylpropyl) phenyl]-hexafluorophosphate(1-)

In accordance with the invention, the monomer stream in which polymericmicrostructures are to be synthesized can incorporate diverse functionalmoieties that produce selected microstructure functionality, foraddressing microstructure applications in, e.g., ambient sensing,self-assembly, rheology, biosensing, drug delivery, and otherapplications. The moieties can be chemically attached in themicrostructure, e.g., by covalent incorporation, or can be physicallyattached or entrapped in the microstructure. Covalently-incorporatedmoieties can be provided as monomers, in the monomer stream, that arepolymerized by the lithography-polymerization step of the microstructuresynthesis. Monomers can be included which alone or in combination withcopolymerized species in a synthesized polymeric microstructure providea selected functionality.

For example, a monomer species can be included in the monomer stream forsynthesizing temperature-sensitive polymer microstructures. The monomerN-isopropylacrylamide, or other suitable monomer, can here be employed.A monomer species can be included in the monomer stream for synthesizingpH-responsive polymer microstructures; monomers such as acrylic acid,methacrylic acid, or other suitable species can here be employed. Amonomer species can further be included in the monomer stream forsynthesizing photosensitive polymer microstructures; a copolymer ofAzobenzene, N,N-Dimethylacrylamide, or other suitable monomer can herebe employed. A monomer species can be included in the monomer stream tosynthesize antigen-responsive polymer microstructures. Here, goatanti-rabbit IgG coupled with N-succinimidylacrylate, or other suitablemonomer, can be employed. With these examples, it is demonstrated that awide range of functionality can be imparted to the synthesizedmicrostructures by the inclusion of selected monomer species.

Further in accordance with the invention, biodegradable monomers, aswell as modified biological material, can be included in the monomerstream. For example, DNA or RNA can be included in the monomer stream.Such can be custom-synthesized or obtained commercially. Polypeptides,antibodies, enzymes, or other such species can also be included in themonomer stream. Each selected species can be modified as-desired toenable covalent incorporation in a polymeric microstructure matrix asthe microstructure is polymerized. Fluorophores and chromophores, suchas fluorescein diacrylate and rhodamine methacrylate, or other moleculesor parts of a larger molecule that can be excited by light to emitfluorescence or selectively absorb light at particular wavelengths, canfurther be included in the monomer stream.

Further in accordance with the invention, selected entities can beprovided in a monomer stream to be physically entrapped in the polymericmatrix of a polymer microstructure as the microstructure is polymerizedby the lithography-polymerization step of the invention. For example,particles such as quantum dots, emulsified liquid droplets, gas bubbles,electrically conductive and metallic particle species, gold or silverfilings or particles, magnetically-sensitive particle species, such asnanometer-sized magnetite or maghemite particles, carbon nanotubes,three-dimensional micromachined or microfabricated structures ofmicroelectronic or other materials, and other species, can be included.If necessary or desirable, these can be physically or chemically linkedto one or more of the monomers provided in the monomer stream. Further,liquid crystals, viruses, whole cells or cellular components, such asmitochondria, proteins, enzymes, nucleic acids, and other such species,can be included in the monomer stream for entrapment in a polymericmicrostructure matrix. Various porogens can be added to the monomerstream to control the porosity of the resulting polymericmicrostructures, such as surfactants, gas bubbles or dissolvablemicroparticles, or nanoparticles such as PMMA nanoparticles.

FIGS. 8A-8C are schematic views of example polymeric microstructuressynthesized in accordance with the invention and including a selectedmoiety. In the example of FIG. 8A, a square mask shape 200 is employedand a selected moiety 205 is incorporated into the polymer matrix of theresulting rectangular microstructure 208. Beads, emulsion droplets,bubbles, cells, particles or other selected entity, such as thosedescribed above, or other species, can be included here.

FIG. 8B is a schematic view of an example polymeric microstructure 210that is also synthesized with a square mask feature 200, but here DNAstrands 212 are incorporated into the polymer matrix of the structure.FIG. 8C is a schematic view of an example polymeric microstructure 214that is synthesized with a circular mask feature 216. The resultingcircular microstructure 214 here includes proteins 218 or other moietyincorporated in the polymer matrix of the microstructure. These examplesare not meant to be limiting but to demonstrate a range of polymericmicrostructures that can be produced with selected species incorporatedinto the microstructure polymer matrix.

The selected one or more species to be added to a monomer forincorporation during the polymeric microstructure synthesis can be mixedwith a monomer by, e.g., vortex mixing, sonication, or other selectedtechnique. Additionally, surfactants can be included with a monomer tostabilize porogens, moieties, or other added entities fromagglomeration. Some species added to a monomer can produce a dispersedphase of particles in the continuous-phase of the fluidic monomers in amonomer stream. In accordance with the invention, such a dispersed phasedoes not constitute that phase which is polymerized to producemicrostructures. As explained above, the continuous phase of the monomerstream is that which is polymerized, with a dispersed phase, if such isnot covalently bonded in the microstructure matrix, being entrapped inthe resulting synthesized polymeric microstructure shapes. The dispersedphase does not in general set the shape or geometry of the synthesizedpolymeric microstructure shapes; it is the lithographic mask shapesimposed on the polymerizing continuous phase that set microstructureshape.

In accordance with the invention, polymeric microstructures can becontrollably synthesized to include two or more functionalities and/orchemistries in an anisotropic arrangement across the plane of amicrostructure. FIGS. 9A and 9C are schematic planar views of twoexample flow configurations for enabling such synthesis. In the exampleflow configuration of FIG. 9A, a Y-shaped flow structure 230 is providedhaving two ports 232, 234, with each port designated to introduce adistinct selected monomer stream 236, 238, respectively, to the crosssectional volume of a microfluidic device like that described above, andhere represented schematically at the region of adjacent alignment 239between the two streams 236, 238.

To synthesize microstructures from the two adjacent monomer streams, aselected mask shape, e.g., the ring shape 240 in FIG. 9A, is employed inthe lithography-polymerization synthesis of the invention describedabove. In a first example synthesis process, the mask 240 and a sourceof illumination are aligned across the interface 241 between the twostreams in a selected manner to incorporate a correspondingly selectedproportion of each stream in the microstructure to be synthesized. In asecond example synthesis process, the mask and source of illuminationare aligned through the two streams, from a side of the streams, as inthe example configuration of lens system 35 of FIG. 1B, to incorporate aproportion of each stream through the thickness of the microstructure.In addition or alternatively, the relative flow rates of the two streamscan be controlled to control the proportion of each stream incorporatedin a microstructure. With these techniques, there is produced apolymeric microstructure that incorporates both monomer streamconstituents in a desired proportion.

FIG. 9B is a schematic view of an example polymeric microstructure 242resulting from a lithography-polymerization step across the two monomerstreams as in FIG. 9A. The microstructure includes a first volume 245corresponding to one of the monomer streams 238 and a second volume 247corresponding to the other monomer stream 236. Although the two volumes245, 247 are shown to be substantially equal in the microstructure ofFIG. 9B, such is not required, and as explained above, any proportioncan be selected. A two-component microstructure like that of FIG. 9B isknown generally as a Janus particle, or two-faced particle. Janusparticles have been demonstrated to be particularly useful for a rangeof applications and the high-through-put, reliable, and reproduciblenature of the polymeric synthesis process of the invention enablesproduction of Janus particles in a practical manner.

Referring to FIG. 9C, in accordance with the invention, the concurrentflowing of three or more monomer streams through a microfluidic devicecan be employed for polymerizing microstructures across the streams togenerate microstructures having many adjacent chemistries and/orfunctionalities. In FIG. 9C there is schematically shown an exampleY-shaped flow structure 250 that accommodates four distinct monomerstreams 252, 254, 256, 258. The streams are all directed through amicrofluidic device, represented here schematically by the adjacentalignment 260 of the streams. A mask shape 262 is provided forlithography and polymerization across the four streams. The resultingpolymeric microstructure 264 is schematically shown in FIG. 9D. Each ofthe four monomer streams 252, 254, 256, 258 produces a correspondingmicrostructure volume 264, 266, 268, 270. The ability to tune theproportion of this number of chemistries in the microstructure enablesgreat flexibility in the design of so-called barcoded microstructuresproviding multiple constituents.

FIGS. 10A-10H are schematic views of additional example polymericmicrostructures that can be synthesized by lithographically defining amicrostructure shape while polymerizing that shape across or through aplurality of concurrently flowing monomer streams. In the example ofFIG. 10A a rectangular mask 300 and four monomer streams are employed toproduce a corresponding rectangular, four-part microstructure 305. Inthe example of FIG. 10B, the rectangular mask 30 is employed with threemonomer streams, one of which includes, e.g., a porogen 205 and one ofwhich includes, e.g., DNA strands 212, in the manner described above, toproduce a three-part microstructure 308 having multiple distinctfunctionalities.

In the example of FIG. 10C a circular mask 310 is employed with sixmonomer streams, three of which are identical and three of which aredistinct, to produce a disc microstructure 311 that incorporates threeregions 312, 314, 316 of identical composition separated by threeregions 318, 320, 322 of distinct composition. In this examplemicrostructure, each of the six resulting regions of the microstructureis characterized by a distinct width, set by the flow rates of thecorresponding monomer streams.

FIG. 10D schematically shows a polymeric microstructure 330 synthesizedacross two monomer streams and employing a mask shape that provides adistinct feature geometry for the microstructure region corresponding toeach of the monomer streams. FIG. 10E schematically shows a polymericmicrostructure 334 synthesized across five monomer streams and employingan S-shaped mask 336. The differentiation in polymer species ispreserved across the structure. In accordance with the invention, theco-flowing monomer streams employed to synthesize polymericmicrostructures can be miscible, chemically similar streams. In thiscase, the interface between different regions in a resultingmicrostructure is not sharp, due to molecular diffusion.Diffusion-limited mixing that is characteristic of laminar flow can beexploited to ensure that the streams flow distinctly through amicrofluidic device, but molecular diffusion between adjacentpolymerized regions can here occur. If co-flowing monomer streamsinstead are immiscible, then a sharp interface between synthesizedpolymeric regions can be enforced, producing microstructures havingregions with sharply-segregated chemistries and differing surfaceenergies.

FIG. 10F is an example of a polymeric microstructure 340 synthesizedacross two monomer streams one of which provides a hydrophilic polymerregion 342 and one of which provides a hydrophobic polymer region 344.The interface 346 between the two regions in the microstructure ischaracteristically curved due to the disparate surface energies of thetwo regions. The amphiphilic nature of such a microstructure can beexploited to enable self-assembly of populations of suchmicrostructures.

FIGS. 10G-10H are schematic views of polymeric microstructures 352, 358,synthesized by lithography and polymerization through multipleco-flowing monomer streams rather than across co-flowing monomerstreams. As shown in FIG. 10G, employing a circular mask 350, athree-material microstructure is produced in which the composition ofthe circular cross section of the structure is changed through thethickness of the structure. Similarly, as shown in FIG. 10H, astar-shaped polymeric microstructure is synthesized across fourdiffering monomer streams to produce a microstructure 358 having adiffering composition through the thickness of the star-structure, withthe composition constant in a given star-shaped plane of the structure.

Example I

A number of microfluidic devices were fabricated by first patterning asilicon substrate with photoresist, SU-8, from Microchem, Newton, Mass.,to define a mold structure corresponding to a rectangular microfluidicdevice cross section as in FIG. 6A. PDMS provided as Sylgard® 184silicon elastomer, from Dow Corning, Midland, Mich., was poured into theresulting positive-relief channels of the silicon substrate to mold anupper PDMS device structure 100 as in FIG. 6A. Rectangular microfluidicdevices of 1 cm in length and three different channel widths weredefined by the patterning, namely, 20 μm-wide, 600 μm-wide, and 1000μm-wide, and three different rectangular microfluidic channel heightswere defined by the patterning, namely, 10 μm in height, 20 μm inheight, and 40 μm in height. A layer of PDMS was spin-coated on glassslides to form bottom surfaces for the microfluidic devices as in FIG.6A. The molded upper PDMS device structures were mated with PDMS-coatedglass slides to form complete microfluidic devices.

For carrying out the lithography-polymerization operations of theinvention with the microfluidic devices, a selected one of the deviceswas mounted on an inverted microscope, the Axiovert 200 invertedmicroscope, from Carl Zeiss MicroImaging, Inc., Thornwood, N.Y. Acharge-coupled device camera (CCD), the Hitachi KP-M1A monochrome CCDcamera, Hitachi America, Ltd., Tarrytown, N.Y., was positioned toacquire images during the microfluidic operations. NIH Image softwarewas employed to capture and process the CCD images.

Dark-field transparency photomasks of selected microstructure shapeswere designed in software with a design tool, AutoCAD® 2005, Autodesk,Inc., San Rafael, Calif., and printed by high-resolution printer by CADArt Services, Poway, Calif. The resulting transparency mask was insertedinto the field stop of the microscope. A source of UV illumination wasprovided by a 100 W OSRAM HBO® mercury short arc lamp, from OSRAMSylvania, Danvers, Mass. A filter set, 11000v2: UV, from ChromaTechnology Corp, Rockingham Vt., allowing wide UV excitation, wasemployed to enable selection of a desired illumination wavelength of 365nm. An electro-programmable shutter system, UniBlitz® VS25, VincentAss., Rochester, N.Y., was driven by a computer-controlled shutterdriver controller, VMM-D1, Vincent Ass., Rochester, N.Y., to producespecified pulses of UV light.

A monomer solution of poly(ethylene glycol)(400) diacrylate (PEG-DA),Polysciences, Warrington, Pa., was employed including a photoinitiatorof 5% (v/v) DAROCUR® 1173, Ciba Specialty Chemicals, Tarrytown, N.Y. Theviscosity of the PEG-DA was reported by the supplier to be 57 cP at 25°C. Additional monomer solutions of trimethylpropane triacrylate,1,6-hexanediol diacrylate, and tri(propylene glycol) diacrylate werealso prepared. A selected one of the solutions was loaded into a syringepump, KDS 100 single-syringe infusion pump, from kdScientific,Holliston, Mass., for delivery through a selected one of themicrofluidic devices.

Pulses of UV light were directed through the transparency mask and themicrofluidic device to a monomer solution passing through themicrofluidic device. As polymeric microstructures were synthesized bythe pulses in the monomer stream, the microstructures were carriedthrough the device in an un-polymerized volume of monomer resulting fromoxidation-induced polymerization inhibition at the device walls. Themonomer-microstructure solution was collected in a reservoir positionedat the output of the device's rectangular channel. The microstructuresin the solution were collected by centrifuge, washed, and thenre-suspended three times in ethanol to dissolve any unpolymerizedmonomer remaining on the microstructures. The microstructures were thenwashed three times in water and then suspended in water.

Four different microscope objectives were employed in separatemicrostructure synthesis processes, namely, 20×, 40×, 63×, and 100×objectives. Table III below provides the actual magnification,theoretical resolution, practical resolution, and depth of field foreach of the objectives.

TABLE III Actual Numerical Theoretical Practical Depth Magni- ApertureResolution Resolution of Field Objective fication (NA) (μm) (μm) (μm)20X 7.8 0.5 0.37 1.29 15.69 40X 15.6 0.75 0.24 0.64 5.38 63X 24.5 1.20.15 0.41 2.13 100X  38.9 1.4 0.13 0.26 1.2

The actual magnification factor is the ratio by which a mask shape sizewas reduced when projected to the monomer stream in a microfluidicdevice. This is different from the magnification of the objectivebecause of an additional 2.57× lens in the optical path between thefield-stop slider and the objective. The theoretic resolution wascalculated using Rayleigh's formula,

${= \frac{0.5\lambda}{NA}},$

where λ is the 365 nm wavelength of illumination employed and NA is thenumerical aperture of the objective. The practical resolution is thesize to which a 10 μm mask feature was reduced using the differentobjectives. The depth of field was calculated using an equation providedby the lens manufacturer, where

${{DOF}({µm})} = {\frac{1000}{7{NA}} + {\frac{\lambda}{2{NA}^{2}}.}}$

This demonstrates that higher magnification objectives provide greaterresolution but lower depth of field.

Analysis of the CCD images of the lithography-polymerization processdetermined that the microstructures were rapidly formed, in less thanabout 0.1 s, due to rapid polymerization kinetics of the selectedmonomers. Oxygen-aided polymerization inhibition near the PDMS surfacesallowed microstructure flow within an unpolymerized monomer streamthrough the complete length of the device. Triangles, squares, hexagons,posts, and other microstructure geometries like that of FIGS. 3-4 weresynthesized. All of the particles showed good fidelity to the originalmask features and had straight sidewalls.

Example II

Square polymeric microstructures were synthesized in the manner ofExample I with square mask features ranging in edge length from 10 μm to500 μm. The microstructure synthesis was duplicated for a 20× objectiveand a 40× objective. The three microfluidic devices described in ExampleI were employed, having channel heights of 10 μm, 20 μm, and 40 μm. ThePEG-DA and DAROCUR® 1173 photoinitiator monomer stream of Example I wasemployed in the microfluidic devices, with the steam flow stopped duringthe lithography-polymerization step, in the stop flow process describedabove. The resulting microstructures were collected and analyzed as inExample I.

The microstructure analysis was conducted to determine the smallest maskfeature that could be polymerized at a given exposure time in thechannels of varying height. FIG. 11A is a plot of the smallest maskfeature that could be polymerized, as a function of exposure time, forthe 20× objective; FIG. 11B is a plot of the smallest mask feature thatcould be polymerized, as a function of exposure time, for the 40×objective. This determination was made by noting when a square of agiven size was polymerized within a tolerance of 10% of a correspondingmask square. As the feature size or the channel height was decreased,longer exposure times were required to polymerize the squaremicrostructures.

Example III

One of the microfluidic devices of Example I having a cross sectionchannel height of 38 μm (intended as 40 μm) was adapted by replacing thePDMS-coated glass slide with an uncoated glass slide, and polymericmicrostructure synthesis was carried out with the monomer as in ExampleI. It was found that the synthesized microstructures stuck to the glassplate. This result is recognized to be due to the absence of the oxygeninhibition effect enabled by the PDMS coating in Example I, therebyallowing the synthesized microstructures to polymerize all the way tothe glass surface.

Polymeric microstructures were synthesized in the manner of Example Iwith a PDMS-coated glass slide device and with an uncoated glass slidedevice, both having a cross-sectional channel height of 38 μm. Anexposure time, t_(exp), of 0.1 s, a 360 μm square mask shape, and a 20×objective were employed with each of the two devices.

Microstructures synthesized with the two microfluidic devices werecollected as in Example I above and analyzed. Microstructuressynthesized with the uncoated glass slide device were characterized by aheight of 35.5 μm. Microstructures synthesized with the PDMS-coatedglass slide device were characterized by a height of 33 μm. Given thatan oxygen-aided inhibition layer was formed during synthesis with thecoated glass slide device, it was determined that the thickness of theunpolymerized lubricating layer was 2.5 μm at both the top and bottomwalls of the device.

This experiment was repeated for rectangular microfluidic device channelheights of 10 μm, 40 μm and 75 μm. For all channel heights, apolymerization inhibition layer thickness of 2.5 μm was measured. Thisresult demonstrates that the inhibition layer thickness is independentof the cross-sectional height of the microfluidic device.

Example IV

A transparency mask was produced as in Example I having features ofdiffering sizes ranging from 10 μm to 500 μm. A fluorescein solution wasemployed as a monomer stream and the lithography-polymerization processof Example I was conducted. Microstructures synthesized by the processwere collected and analyzed in the manner of Example I.

It was found that the light passing through the transparency mask in thefield-stop plane of the microscope objective had a featuresize-dependent intensity below a critical feature size of 250 μm. Abovethis critical feature size, the light intensity was equal through anymask feature dimensions and led to the polymerization of all suchfeatures in equal exposure durations. Below this critical size, the beamintensity was found to decrease with the size of the feature. Thismeasured variation in intensity is recognized to be caused by clippingof the light passing through the mask, resulting from the increasingdivergence of the light beam as the size of the aperture was decreased.

Example V

The lithography-polymerization process of Example I was conducted withthe microfluidic devices having rectangular channel heights of 10 μm, 20μm, and 40 μm, and employing the 20× objective and the PEG-DA andphotoinitiator monomer stream. Four different polymerization processeswere carried out, with the velocity of the monomer stream varied betweenabout 100 μm/s and about 1700 μm/s between the processes. Polymerizedmicrostructures synthesized by the processes were collected and analyzedas in Example I.

FIG. 12 is a plot of the minimum measured mask feature size that couldbe polymerized, with a tolerance factor of 10% in length scale, at agiven velocity of the PEG-DA monomer stream. This data can be employedas a guide to determine the maximum monomer stream velocity that can beemployed to synthesize a selected mask feature size. Lower monomerstream velocities were required to generate microstructures havingsmaller features, while larger microstructures could be synthesized athigher monomer stream velocities.

Example VI

The lithography-polymerization process of Example I was conducted with atwo-port flow structure fabricated in the manner of the one-portstructure of Example I, but here employing a Y-shaped geometry like thatof FIG. 9A, for producing rectangular Janus particles. Thecross-sectional channel region in which the monomer streams wereco-flowing was of a height of 200 μm and a width of 20 μm. In a firstport of the structure was directed a flow of the PEG-DA and DAROCUR®1173 monomer stream of Example I. In the second port of the structurewas directed a flow of DAROCUR® 1173 and PEG-DA with arhodamine-labelled crosslinker, a 0.005 wt % solution of the fluorescentmethacryloxyethyl thiocarbamoyl rhodamine B, from Polysciences, Inc.,Warrington, Pa., to fluorescently label the polymer. A rectangular maskfeature was aligned across the flow of the two monomer streams, and thelithography-polymerization was carried out as in Example I, with thepolymerized microstructures collected and analyzed. It was verified byfluorescence microscopy that the fluorescently labeled and thenon-labeled sections were distinctly formed as in FIG. 9B.

Example VII

The lithography-polymerization process of Example VI, with theexperimental conditions of Example I, was conducted with two-port PDMSY-shaped microfluidic devices having a rectangular cross sectionalchannel width of 200 μm and 300 μm, and a height of 30 μm. A reservoirwas cut in each of the PDMS microfluidic devices to collect thesynthesized microstructures. The shutter control was set to provide alithographic exposure time of 0.03 s; with a pause of 5 s betweensuccessive exposures.

Amphiphilic polymeric microstructures were synthesized with ahydrophobic phase monomer stream consisting of a 5% (v/v) DAROCUR® 1173,photoinitiator in tri(methylpropane) triacrylate, (TMPTA) PolysciencesInc., Warrington, Pa. A hydrophilic phase monomer stream was providedconsisting of 5% (v/v) solutions of the DAROCUR® 1173 photoinitiator in65% aqueous solution of poly(ethylene glycol)(600) (PEG-DA). BecauseTMPTA is insoluble in water, the two monomer streams were immiscible.Both PEG-DA and TMPTA are reported by the supplier to have viscositiesof 90 cP and 106 cP, respectively, at 25° C.

The two monomer streams were controlled with a syringe pump to a flowvelocity between about 100 and about 300 μm/s, constraining the flowregime to one in which parallel co-flow of the two streams was enforced.Because the two monomer streams were immiscible, they cooed all the wayto the exit of the microfluidic device. This led to a segregation of thetwo phases all along the interface and was convenient for the formationof large numbers of microstructures where diffusive mixing mightotherwise have constrained the region available for polymerization.

Transparency masks were produced in the manner of Example I withmicrostructure wedge shapes that fall in the spectrum of shapes betweentriangles and rectangles. Such are the two-dimensional analogs ofamphiphilic molecules as represented by objects in the spectrum ofshapes between a cone and a cylinder, where the body of the objectrepresents the hydrophobic tail and the hydrophilic heat is representedby a circular face. This spectrum of shapes is used to show the effectof geometry on packing.

FIG. 13 is a schematic representation of the lithography-polymerizationprocess with this mask including a row of 5 wedge shapes for projectionto the two flowing monomer streams, such that wedge-shapedmicrostructures were polymerized, five at a time, across the co-flowingstreams. A 20× microscope objective was employed, resulting inmicrostructures having that were approximately ⅛th of their mask size.Copolymerization of the two acrylates used led to a chemical linkage ofthe hydrophilic and hydrophobic sections at their interface, lendingstability to the microstructures formed. As thelithography-polymerization process proceeded, wedge-shapedmicrostructures were formed and flowed continuously through themicrofluidic device in unpolymerized monomer flow resulting fromoxygen-induced inhibition of polymerization at PDMS surfaces in themanner described above.

The microstructures were collected in unpolymerized monomer in themanner described above and then dispersed in ethanol in order to avoidclustering. Because both the hydrophilic and hydrophobic polymericprecursors were completely soluble in ethanol, there was no preferentialalignment of the microstructures in the ethanol. But the hydrophilicportion of each microstructure was found to swell to a greater extent inethanol, leading to a slight distortion in the original shape. When dry,the hydrophilic portion of each microstructure was found to shrink incomparison to the hydrophobic phase.

The sizes of the synthesized microstructures were estimated as they werebeing formed by capturing images of the flowing microstructures a lowcamera exposure time of 1/10000 s. Statistics were estimated on a 100consecutive microstructures formed at an exposure time of 0.03 s. Acomprehensive analysis of particle monodispersity was performed bymeasuring the distribution of the five different length variables of themicrostructure wedge shape, w₁, w₂, w₃, h₁ and h₂ as given in FIG. 14.In addition to the mask-defined edges of the microstructures, w₁, w₃,and (h₁+h₂), the lengths of their hydrophilic portion, h₁ andhydrophobic portion h₂ are recognized to be dictated by the preciseposition of the interface when polymerization occurs. These lengths wereall measured as the microstructure formed in the microfluidic device.

FIGS. 15A-15E are histogram plots of the measured distributions of thesedimensions. In all five dimensions that were measured, the coefficientof variation (COV) in size was less than 2.5% so that the particles canbe correctly classified as monodisperse, wherein >90% of thedistribution was within 5% of the median microstructure size. It wasfurther found that by changing the length of the hydrophilic portion h₁and the hydrophobic portion, h₂, the extent of a microstructure'samphiphilicity could also be tightly controlled. The thickness of theparticles, in the y direction in FIG. 14, was measured to be 25 μm,which corresponds to an unpolymerized lubrication layer of thickness 2.5μm at the top and bottom microfluidic device walls, as in the examplesabove.

An inspection of the cross section of one of the wedge microstructuresindicated that the interface between the hydrophilic portion and thehydrophobic portion of the microstructure was characterized by a finitecurvature, as in FIG. 10F, and here defined in FIG. 16. This curvaturewas caused by the immiscibility of the two flowing monomer streams,leading to a sharp, curved interface The hydrophobic (‘o’) phasepreferentially wet the PDMS over the hydrophilic (‘w’) phase. Thecontact angle at the interface between the two phases and the PDMS wall,β, was dictated by the interfacial tension between the hydrophilic phaseand the hydrophobic phase as well as by the solid-liquid interfacialtension between the PDMS and the two phases.

Using simple geometry, the radius of curvature, R, at the interface canbe given as R=H/2 cos β, where H is the height of the microfluidicdevice cross section. For the experimental parameters given above, apredicted radius of 17.4 μm was determined this was found to match wellwith the experimentally determined value of 16±1.5 μm. This demonstratesthat the interfacial properties of the two monomer streams can be usedto tune the curvature of the interface between the two microstructuresections.

The extent of cross-linking of the particles in the two different phaseswas characterized by measuring the percentage of double bonds convertedusing FTIR spectroscopy performed on a Nikolet spectrometer, from ThermoElectron Corp., Waltham, Mass., by measuring the decrease in terminalC═C stretch at 1635 cm⁻¹ in a thin film of cross-linked polymer. Toperform FTIR, individual samples of either the hydrophilic or thehydrophobic monomer were prepared by loading the respective oligomerinto a channel with the same dimensions as the microfluidic device usedand then given exposure doses of 0.03 s, as with the synthesizedmicrostructures, and 120 s, for fully cross-linked. Strips of polymerfilm were formed that were used for FTIR measurements. Contact angle andsurface tension measurements were performed using a DSA 10 tensiometer,from Druss USA, Matthews, N.C.

FIG. 17A is a plot of the FTIR spectrum for the hydrophobic monomerstream and FIG. 17B is a plot of the FTIR spectrum for the hydrophilicmonomer stream, both for exposure times of 0 s, 0.03 s, and 120 s. Inthe 0.03 s duration of exposure to UV light, the conversion of doublebonds was found to be 47% in the hydrophilic phase and 35% in thehydrophobic phase. Because the conversion of double bonds required tocross-link multifunctional acrylates successfully is typically less than5%, it is concluded that this exposure dose is sufficient to cross-linkthe microstructures.

Like amphiphilic molecules, microstructures possessing both hydrophilicand hydrophobic sections exhibit a tendency to orient themselves inorder to minimize their surface energy. While thermal energy alone isinsufficient to enable these microstructure to explore their energylandscape, external energy provided by, e.g., agitation was employed toaid the microstructures to find their energy minima and self-assemble.The wedge-shaped amphiphilic microstructures were isolated and induced,using agitation, to assemble either in a pure aqueous phase or at theinterface of w/o or o/w emulsions. The results showed that the particleshave a strong tendency to orient themselves in order to minimize theirsurface energy.

This method of immiscible microstructure synthesis is of sufficientgenerality to enable synthesis of a wide range of non-sphericalparticles with such chemical anisotropy; e.g., amphiphilic particleswith a rod-like hydrophobic tail and a disk-shaped hydrophilic head canbe synthesized. Such a library of particles can be useful when studyingthe effect of geometry and chemical anisotropy on meso-scale selfassembly and rheology. In addition, structures with more complicatedmotifs like w-o-w can also be formed quite easily.

With these examples and the description above, it is demonstrated thatthe invention provides lithographic-based microfluidic methodology thatcan be employed to continuously or near-continuously synthesizepolymeric microstructures of varied complex shapes, chemistries, andfunctionalities with an elegant dual lithography-polymerization step.The morphology and chemistry of the microstructures that are synthesizedcan be independently controlled to produce large numbers of uniquelyshaped, functionalized polymeric microstructures for applicationsincluding drug delivery, biosensing, microactuation, and fundamentalstudies on self-assembly and rheology, among others. Thehigh-through-put of the synthesis processes of the invention enables thepractical achievement of polymeric microstructure synthesis on a scalethat is required for many of these applications.

It is recognized, of course, that those skilled in the art may makevarious modifications and additions to the embodiments described abovewithout departing from the spirit and scope of the present contributionto the art. Accordingly, it is to be understood that the protectionsought to be afforded hereby should be deemed to extend to the subjectmatter claims and all equivalents thereof fairly within the scope of theinvention.

1. A non-spheroidal polymeric microstructure having a plurality ofdistinct material regions.
 2. The polymeric microstructure of claim 1provided in a population of monodisperse non-spheroidal polymericmicrostructures.
 3. A planar polymeric microstructure having a pluralityof distinct material regions.
 4. The polymeric microstructure of claim 3wherein the polymeric microstructure is characterized by an anisotropicmaterial composition.
 5. The polymeric microstructure of claim 3 whereinat least two of the distinct regions are of a distinct polymericcomposition.
 6. The polymeric microstructure of claim 3 wherein at leastone of the distinct regions includes a porogen.
 7. The polymericmicrostructure of claim 3 wherein at least one of the distinct regionsincludes a covalently bonded moiety.
 8. The polymeric microstructure ofclaim 3 wherein at least one of the distinct regions is hydrophobic andat least one of the distinct regions is hydrophilic.
 9. The polymericmicrostructure of claim 3 further comprising three-dimensional features.10. The polymeric microstructure of claim 3 further comprisingtwo-dimensional extruded features.
 11. The polymeric microstructure ofclaim 3 further comprising at least one straight edge.
 12. The polymericmicrostructure of claim 11 further comprising a plurality of straightedges.
 13. The polymeric microstructure of claim 3 further comprising atleast one rounded feature.
 14. The polymeric microstructure of claim 3wherein the microstructure is hollow.
 15. The polymeric microstructureof claim 3 wherein the microstructure is cylindrical.
 16. The polymericmicrostructure of claim 3 wherein the microstructure is non-continuousacross a plane of the microstructure.
 17. The polymeric microstructureof claim 16 wherein apertures in the microstructure result in thenon-continuity of the microstructure.
 18. The polymeric microstructureof claim 17 wherein at least one aperture is straight-edged.
 19. Thepolymeric microstructure of claim 17 wherein at least one aperture isrounded.
 20. The polymeric microstructure of claim 3 wherein themicrostructure is characterized by a sensitivity to temperature.
 21. Thepolymeric microstructure of claim 3 wherein the microstructure ischaracterized by a sensitivity to pH.
 22. The polymeric microstructureof claim 3 wherein the microstructure is characterized by a sensitivityto illumination.
 23. The polymeric microstructure of claim 3 wherein themicrostructure is characterized by a responsiveness to at least oneantigen.
 24. The polymeric microstructure of claim 3 wherein themicrostructure is biodegradable.
 25. The polymeric microstructure ofclaim 3 wherein the microstructure includes biological material.
 26. Thepolymeric microstructure of claim 25 wherein the biological materialcomprises DNA.
 27. The polymeric microstructure of claim 25 wherein thebiological material comprises RNA.
 28. The polymeric microstructure ofclaim 25 wherein the biological material comprises a polypeptide. 29.The polymeric microstructure of claim 25 wherein the biological materialcomprises an enzyme.
 30. The polymeric microstructure of claim 25wherein the biological material comprises an antibody.
 31. The polymericmicrostructure of claim 25 wherein the biological material comprises atleast a portion of a cell.
 32. The polymeric microstructure of claim 25wherein the biological material comprises mitochondria.
 33. Thepolymeric microstructure of claim 3 wherein the microstructure includesa fluorophore.
 34. The polymeric microstructure of claim 3 wherein themicrostructure includes a chromophore.
 35. The polymeric microstructureof claim 3 wherein the microstructure includes a porogen.
 36. Thepolymeric microstructure of claim 3 wherein the microstructure includesparticles.
 37. The polymeric microstructure of 3 wherein themicrostructure includes quantum dots.
 38. The polymeric microstructureof claim 3 wherein the microstructure includes carbon nanotubes.
 39. Thepolymeric microstructure of claim 3 wherein the microstructure includesemulsified liquid droplets.
 40. The polymeric microstructure of claim 3wherein the microstructure includes gas bubbles.
 41. The polymericmicrostructure of claim 3 wherein the microstructure includeselectrically conducting particles.
 42. The polymeric microstructure ofclaim 3 wherein the microstructure includes magnetizable particles. 43.The polymeric microstructure of claim 3 wherein the microstructureincludes three-dimensional structures.
 44. The polymeric microstructureof claim 3 wherein the microstructure includes liquid crystals.
 45. Thepolymeric microstructure of claim 3 wherein the microstructure comprisesa microstructure chain in which microstructures are linked to the chainby an overlapping polymerized region.
 46. The polymeric microstructureof claim 3 provided in a population of monodisperse polymericmicrostructures.